Centro de Biología Molecular 'Severo Ochoa' (Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid), c/ Nicolás Cabrera 1, Campus de Cantoblanco-UAM, 28049 Madrid, Spain.
Centro de Biología Molecular 'Severo Ochoa' (Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid), c/ Nicolás Cabrera 1, Campus de Cantoblanco-UAM, 28049 Madrid, Spain.
Trends Biotechnol. 2021 Feb;39(2):194-210. doi: 10.1016/j.tibtech.2020.06.008. Epub 2020 Jul 8.
Reverse transcriptases (RTs) are enzymes that can generate a complementary strand of DNA (cDNA) from RNA. Coupled with PCR, RTs have been widely used to detect RNAs and to clone expressed genes. Classical retroviral RTs have been improved by protein engineering. These enzymes and newly characterized RTs are key elements in the development of next-generation sequencing techniques that are now being applied to the study of transcriptomics. In addition, engineered RTs fused to a CRISPR/Cas9 nickase have recently shown great potential as tools to manipulate eukaryotic genomes. In this review, we discuss the properties and uses of wild type and engineered RTs in biotechnological applications, from conventional RT-PCR to recently introduced prime editing.
逆转录酶（RTs）是一种能够从 RNA 生成互补 DNA（cDNA）链的酶。与 PCR 结合使用，RT 已被广泛用于检测 RNA 并克隆表达基因。经典的逆转录病毒 RT 通过蛋白质工程得到了改进。这些酶和新鉴定的 RT 是开发新一代测序技术的关键要素，该技术现在正被应用于转录组学的研究。此外，最近与 CRISPR/Cas9 切口酶融合的工程 RT 已显示出作为工具来操纵真核生物基因组的巨大潜力。在这篇综述中，我们讨论了野生型和工程 RT 在生物技术应用中的性质和用途，从传统的 RT-PCR 到最近引入的 Prime 编辑。
